This paper reviews recent development and applications of high-speed countercurrent chromatography (HSCCC) in the separations on the analytical scale,the semi-preparative and preparative scale of natural products,antibiotics,proteins,inorganic compounds etc.Several series of two-phase solvent systems to facilitate the systematic search suitable for HSCCC are introduced.And the new countercurrent chromatography (CCC)technologies such as HSCCC coupled with MS,pH-zone-refining countercurrent chromatography,and ion-pairing Countercurrent Chromatography are explained and prospected in term of applications.

AIM: To optimize the excipient formulation of lactose,HPMC_(SH4000) and Carbopol 71 G in breviscapin sustained release tablets by mixture uniform design. METHODS: Different formulations,single factor f_2 and multifactors of cumulative release as index,were evaluated respectively. RESULTS: The optical formulations obtained by the two methods were identical,and the tablets with optical formulation had ideal sustained release. CONCLUSION: Mixture uniform design is simple,direct and uniform.It can be used in optimization of formulation with invariable amount.

AIM:To establish the fingerprint analysis method of Yaotongning Capsule by HPLC-DAD.METHODS:This experiment used homogeneous design to optimize extract method.the analysis was performed on an Agilent C_ 18(4.6 mm?250 mm,5 ?m.)column with a acetonitrile-0.2 acetic acid,0.2% triethylamine.Detection time was 60 min.The flow rate was 1.0 mL/min and the wavelength was at 254 nm.Effect of solvents time and method of extract on experiment was studied,then calculated fingerprint similarities to evaluate,the samples' stability.RESULTS:15 peaks including strychnine,brucine,glycyrrhizic acid,ecdy sterone and ephephedrine,were separated on HPLC fingerprint in Yaotongning Capsule.CONCLUSION:The method is reliable,accurate and can be used as a quality control for Yaotongning Capsule.

Objective: To introduce a new way, which facilitates the systematic search and evaluation of the solvent systems suitable for high-speed countercurrent chromatography (HSCCC). Methods: The software integrated with the thermodynamics solution functions and theory is directive to calculate the composition, physical parameters, such as volume, viscosity, dielectric constants, and polarity parameters of the upper and lower phases. Results: Separation of Guizhi-decoction Fr. A by HSCCC, one of traditional Chinese medcines, is conducted to prove the applicability of this software in selection and optimization of solvent systems. Conclusion: This software can be used to select and optimize the solvent systems of HSCCC. It is supposed to extend this method.

BACKGROUND:Diabetic cystopathy is one of the most common chronic diabetic complications. The establishment of animal models of diabetic cystopathy wil provide experimental animal platform for relevant research.
OBJECTIVE:To establish a guinea pig model of diabetic cystopathy and to evaluate its urodynamic characteristics.
METHODS:Fifty short-hair Britain female guinea pigs were randomly divided into two groups, 42 as the experiment group and the other 8 as the control group. The experiment group was intraperitoneal y injected with streptozotocin to induce diabetes. The control group received injection of blank citric acid buffered solution. Diabetic guinea pigs were detected by urinary dynamics test at 9 and 12 weeks. Diabetic guinea pigs were further assigned into diabetic cystopathy subgroup and compensated subgroup. The urodynamic parameters of three groups were compared.
RESULTS AND CONCLUSION:Twenty of 42 guinea pigs were successful y induced diabetes by the injection of streptozotocin. At 9 weeks after the injection, bladder function compensation was present in six diabetic guinea pigs while bladder function was decompensated in another three diabetic guinea pigs. At 12 weeks, bladder function compensation was present in one diabetic guinea pig, while another eight guinea pigs were confirmed with diabetic cystopathy (88.89%). In the diabetic cystopathy subgroup, the residual urine volume was increased (0.72±0.08) mL, maximal detrusor pressure was decreased (0.63±0.05) kPa, maximum bladder capacity was increased (2.01±0.05) mL, and bladder compliance was increased (0.34±0.04) mL/kPa. There were significant differences compared with the compensated subgroup and the control group (P<0.001). Diabetic cystopathy occurs at 12 weeks after diabetic models are successful y established in guinea pigs, and urodynamic changes are mainly the increase of residual urine volume.

Objective To investigate the expression of Semaphorin 3F (SEMA3F) protein in hepatocellular carcinoma (HCC) and to demonstrate its relationship with clinicopathological features and prognosis of HCC. Methods Western Blotting was carried out in 32 hepatocellular carcinoma samples and matched perineoplastic tissues to detect the expression of SEMA3F protein. The relationship between SEMA3F protein expression and clinicopathological features as well as prognosis of HCC patients was analyzed. Immunohistochemistry was used to show the location of SEMA3F in HCC cells and its relationship with microvessel density (MVD). Results The expression of SEMA3F protein in HCC tissues was significantly higher than in the perineoplastic tissues (447.78± 48.26 vs 618.93 ±61.23, P＜0. 05) and it was correlated closely with tumor capsulation and tumor nodular number (P＜0.05). Based on the Western Blotting and clinical follow-up data, we found that the survival time of HCC patients with a higher SEMA3F expression level was longer than those with a lower level, and the recurrent/metastatic time of HCC patients was significantly different between these two groups (P＜0.01). Immunohistochemistry demonstrated that SEMA3F protein localized in the cytoplasm of HCC cells and its expression correlated with HCC MVD. MVD in the low-level group was higher than the high-level group (115.6±30.38 vs 86. 56±17.94, P＜0.01). Conclusions SEMA3F expression in HCC was significantly down-regulated and correlated closely with tumor-capsulation, nodular number, and MVD, implicating SEMA3F may play an important role in recurrence and metastasis of HCC. It can be regarded as a prognostic marker in HCC patients.

Objective To determine the serum levels of cross-linked telopoptide of type Ⅰ collagen (ICTP),alkaline phosphatase (ALP) in patients with primary malignant bone tumor,primary benign bone tumor and malignant tumor metastasized to the bone,and to explore the clinical value of ICTP and ALP in identification and diagnosis of bone tumor.Methods Sixteen primary malignant bone tumor patients,16 primary benign bone tumor patients and 18 malignant tumor metastasized to the bone patients in 2012 were studied.Serum ALP was assayed by SFBC rate method and ICTP by enzyme linked immunosorbent assay (EIA).Results The serum levels of ICTP was not significantly different between primary benign bone tumors and normal control group (P ＞ 0.05),but the other between-groups had statistically significant difference (P ＜ 0.05).The serum levels of ALP in malignant tumor metastasized to the bone was significantly higher than the rest of the group (P ＜0.01),but the difference between the remaining groups was not statistically significant (P ＞ 0.05).The area under roc curve (AUC) of ICTP for diagnosis of primary benign bone tumors,malignant tumor metastasized to the bone,primary malignant bone tumor (0.923,0.926,0.874) was higher than the ALP (0.354,0.702,0.865).Conclusions Serum ICTP and ALP were sensitive and convenient biochemical indices which reflected metabolism of patients with bone tumor.Serum ICTP was more specific and sensitive than ALP and they have clinical importance for differential diagnosis as an index of bone tumor.

The objective of this study is to fully investigate the therapeutic effect and mechanisms of action of Gegen Qinlian decoction (GD) on type 2 diabetes mellitus (type 2 DM). A rat model of type 2 DM was established with the combination of high-fat diet and multiple low doses of streptozotocin (STZ). Biochemical indicators related to glucose metabolism disorders, insulin resistance, oxidative stress were observed. The type 2 DM rats were administrated with GD for 80 days, the above-mentioned indexes were detected. The results indicated that the hepatic glycogen synthesis level was promoted, fasting blood glucose level and fasting blood insulin level were significantly reduced, insulin sensitivity index was significantly improved; the level of superoxide dismutase (SOD) was increased and the level of malondialdehyde (MDA) was reduced; pathologic morphology of pancreas and kidney was ameliorated in the GD group. It was indicated that the therapeutic mechanisms of action of GD on type 2 DM might be related to its effect of ameliorating glucose metabolism disorders, relieving insulin resistance, increasing the tissues' sensitivity to insulin, improving the antioxidative ability of living system, GD has therapeutic effect on type 2 DM and protective effects against damaged pancreatic function.

Objective: To establish two methods (HPLC UV and HPLC ELSD)for the determination of 3 kinds of cholic acids in Qingkailing Injection, and compare them with UV method. Methods: Two HPLC methods detected by UV and ELSD, respectively have been developed: Phenomenex Luna C 18 column(250mm?4.6mm,5?)with linear gradient elution using methanol acetotrile(4∶1) and 0.1% aqueous formic acid. Results: Three kinds of cholic acids in Qingkailing could be determined simultaneously by HPLC UV and HPLC ELSD. The recovery of three method were 100%,97%,95%～96% for HPLC/ELSD,HPLC/UV,UV, respectively. Conclusion: The HPLC ELSD method was accurate and specific to determine the content of cholic acids in Qingkailing Injection.

Objective To investigate the effects of antidepressant therapy on whole regulating function of OPG/RANKL/RANK system in ovariectomized (OVX) rat model with depressive disorder.Methods Forty sexually mature female SD rats were used as subjects.The rats were randomly divided into 8 groups as following:sham group(Sn),depression group(Dn),the OVX group (On),the OVX with depression group(ODn),OVX with depression +sertraline group (ODs),and OVX with depression + citalopram (ODx),OVX with depression + reboxetine group(ODr),OVX with depression + venlafaxine group (ODw).Bilateral oophorectomy was applied to establish the ovariectomized rats model.Chronic unpredictable mild stress(CUMS) was used to estalish depression model.Open-field test and sugar consumption experiment were applied before the experiment and the 28th day of the experiment.Immunohistochemistry and Quantitative Real-time PCR were used to observe the expression of OPG,RANKL and RANK.Results After chronic unpredictable mild stress the scores of horizontal motion,vertical motion and percentage of sugar consumption experiment in Dn group were decreased contrast to the in control Sn group (P＜0.05)and the scores in ODn group were decreased in contrast to the On group (P＜0.05).The protein and gene expression of RANK in Dn group and On group were increased in contrast to the Sn group (P＜0.05).The protein and gene expression of RANKL in Dn group and On group were increased in contrast to the Sn group (P＜0.05).The gene expression of OPG in ODs group(5.94±0.79),ODw group(5.99± 1.08) and ODr group(6.59±0.69) was decreased in contrast to the ODn group (4.96±0.51) (P＜0.05).The gene expression of RANK in ODs group (2.82±0.23),ODx group(3.03±0.04) was increased in contrast to the ODn group(3.44±0.18) (P＜0.05).The gene expression of RANKL in ODs group(4.82±0.94),ODx group(5.00±0.82) and ODw group(5.23±0.88)was decreased in contrast to the ODn group (3.69± 0.69) (P＜0.05).Conclusion The expression of RANK and RANKL in femoral neck is increased both in depressive disorder model rats and ovariectomized model rats.The antidepressant can up-regulate the expression of RANK,and down-regulate the expression of OPG and RANKL in the OVX with depression group.Of the effect,sertraline is the strongest,however reboxetine mesylate is the poorest.